[Detection of streptomycin resistance in Mycobacterium tuberculosis clinical isolates by denaturing high-performance liquid chromatography and DNA sequencing].
OBJECTIVE
To determine the rpsL and rrs gene mutation in Mycobacterium tuberculosis (M. tuberculosis) and compare the consistency between the results of denaturing high-performance liquid chromatography (DHPLC) and those of DNA sequencing.


METHODS
The values of streptomycin minimum inhibitory concentration (MIC) against 215 M. tuberculosis clinical isolates, 115 being streptomycin-resistant and 100 being susceptible by a routine proportional method, were tested by DHPLC. DNA sequencing was conducted to detect the rpsL and rrs mutation.


RESULTS
98 of the 115 streptomycin-resistant isolates (85.2%) harbored rpsL and/or rrs mutation, 76.5% of which being rpsL mutation (88/115). There was no significant correlation between the MIC values and mutation types. No mutation was found in all the susceptible isolates. There was a complete consistency between the DHPLC results and those of DNA sequencing.


CONCLUSIONS
DHPLC can be regarded as a useful and powerful tool to detect the streptomycin resistance detection in M. tuberculosis.
Shi
Rui-ru
RR
Central Laboratory of Henan Provincial Chest Hospital, Zhengzhou 450003, China. shiruiru@hotmail.com
Zhang
Jian-yuan
JY
Yuan
Xue-qin
XQ
Sun
Zhao-gang
ZG
Li
Chuan-you
CY
D000293:Adolescent; D000328:Adult; D000368:Aged; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D024881:Drug Resistance, Bacterial; D006801:Humans; D008826:Microbial Sensitivity Tests; D008875:Middle Aged; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D015203:Reproducibility of Results; D012269:Ribosomal Proteins / Q000235:genetics; D013307:Streptomycin / Q000494:pharmacology*; D014376:Tuberculosis / Q000382:microbiology; D055815:Young Adult
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012269:Ribosomal Proteins; D013307:Streptomycin